1. Field of the Invention
The invention relates to electrophoresis and, more particularly, to new gel trays and membranes pre-imppregnated with developing reagents, which can be used in carrying out new clinical and medico-legal processes that have been rendered practical by recently developed microelectrophoretic equipment.
2. The Prior Art
In a copending patent application by the present inventor, Ser. No. 744,574, filed on Nov. 24, 1976, now U.S. Pat. No. 4,061,561 there are disclosed several improvements of microelectrophoretic apparatus which greatly simplify and standardize the microelectrophoretic process and which include: (a) designs that allow the use of either a membrane or a tray with the same basic apparatus; (b) designs which allow the two-dimensional resolution of a protein sample placed on a gel-filled tray, (c) a sample applicator and accessory equipment which permit the simultaneous and precise positioning of e.g., 10 samples.
This apparatus can be used advantageously with various conventional devices and techniques which include comartmentalized gel trays, as disclosed by Siebert et al. in U.S. Pat. No. 3,616,387 (FIG. 5 and column 3, lines 53 to 55) and slotted membranes, as disclosed by Zec in U.S. Pat. No. 3,317,418 (FIG. 7). Also, when used in conjunction with the further refined articles that are disclosed in the present application, it makes available to research scientists and laboratory technicians new and improved methodology for separation of specific proteins from microliter quantities of blood. These specific proteins are then identified through use of immunologic techniques or, more commonly, through use of indicator dyes which chemically unite with one specific type of protein and no other. These dyes make contact with the electrophoresed samples by means of an overlay, eithr cellulose acetate membrane upon gel, or gel upon cellulose acetate membrane. The overlays have been pre-impregnated with the appropriate specific substrate so that a permanent visual record of the pattern is produced on the cellulose acetate membrane, whether the membrane is serving as the supporting medium or as the overlay. This technique is described by Grunbaum in "An Automatic One-to-Eight Sample Applicator for Fast Qualitative and Quantitative Microelectrophoresis of Plasma Proteins . . . ," Microchem. J. 20, 495-510 (1975).
This apparatus is useful in research and application in the fields of Medicine, Immunology, Genetics, Biochemistry, and Forensic Science. Electrophoretic procedures which greatly increase in utility with this apparatus include determinations of significant polymorphic enzyme systems such as lactic acid dehydrogenase (LDH), alkaline phosphatase (AP), and creatine phosphokinase (CPK). It can be used for diagnostic purposes through determination of specific antibodies or antigens in the blood. In the forensic laboratory, it can be used in the phenotyping of genetic variants of enzymes and other proteins in blood for the purose of identification or individualization.
A partial list of the factors in blood that can be determined using this apparatus include the following:
lactic acid dehydrogenase (LDH) PA1 alkaline phosphatase (AP) PA1 creatine phosphokinase (CPK) PA1 erythrocyte acid phosphatase (EAP) PA1 glucose-6-phosphate dehydrogenase (G-6PD) PA1 adenylate kinase (AK) PA1 hemoglobin (Hb) PA1 haptoglobin (Hp) PA1 group specific component (Gc) PA1 lipoprotein (Lp) PA1 adenosine deaminase (ADA) PA1 6-phosphogluconate dehydrogenase (6-PGD) PA1 Glyoxylase I (GLO-I) PA1 glutamic pyruvic transaminase (GPT) PA1 esterase D (EsD) PA1 Glutathione reductese (GsR) PA1 Immunoglobulins (Ig)
Methodology for the phenotyping of additional systems is being developed.